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Matrigel coated boyden transwell chambers

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Matrigel-coated Boyden transwell chambers are a cell migration and invasion assay used to measure the ability of cells to migrate through a porous membrane. The chambers are coated with Matrigel, a basement membrane extract, to simulate the extracellular matrix. Cells are seeded in the upper chamber, and the migration of cells through the membrane is measured over time.

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3 protocols using matrigel coated boyden transwell chambers

1

Matrigel Invasion Assay Protocol

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In vitro invasion assay was done using matrigel-coated Boyden transwell chambers from BD Biosciences. Transwells were soaked in 50 µl serum-free medium for 30 minutes at RT and then ~40,000 cells (in RPMI medium containing 0.1% BSA) were placed into the upper chambers. These chambers were then placed in 24 well plate (one per well) on top of 500 µl RPMI medium containing 3% fetal bovine serum as chemo-attractant. Inhibitors were added directly to the medium and mixed. Then the cells were incubated at 37 °C in the CO2 incubator for 16 hours. Non-invaded cells along with matrigel in the upper chambers were scraped with a cotton tipped applicator and then the membranes were fixed in methanol, stained with 0.025% crystal violet, and observed under a Leica microscope at x200.
Monga J., Subramani D., Bharathan A, & Ghosh J. (2020). Pharmacological and genetic targeting of 5-lipoxygenase interrupts c-Myc oncogenic signaling and kills enzalutamide-resistant prostate cancer cells via apoptosis. Scientific Reports, 10, 6649.
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2

Transwell Invasion Assay Protocol

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Invasion assay was done in matrigel-coated Boyden transwell chambers from BD Biosciences. Transwell chambers were soaked in 50 µl serum-free RPMI medium for 30 minutes at RT and then ~40,000 cells (in RPMI medium containing 0.1% BSA) were placed into the upper chambers with or without drugs. These chambers were then placed in 24 well plate (one per well) on top of 500 µl RPMI medium containing 3% fetal serum as chemo-attractant. Inhibitors were added directly to the medium and mixed. Then the cells were incubated at 37°C in the CO2 incubator for 16 hours. Non-invaded cells along with matrigel in the upper chambers were scraped with a cotton tip applicator and then the membranes were fixed in methanol, stained with 0.25% crystal violet, and observed under a Leica microscope at ×200.
Sarveswaran S., Ghosh R., Parikh R, & Ghosh J. (2016). Wedelolactone, an anti-inflammatory botanical, interrupts c-Myc oncogenic signaling and synergizes with enzalutamide to induce apoptosis in prostate cancer cells. Molecular cancer therapeutics, 15(11), 2791-2801.
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3

Matrigel-based Invasion Assay

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In vitro invasion assay was done using Matrigel-coated Boyden transwell chambers from BD Biosciences. Transwells were soaked in 50-mL serum-free medium for 30 minutes at room temperature and then $40,000 cells (in RPMI medium containing 0.1% BSA) were placed into the upper chambers. These chambers were then placed in 24 well plate (one per well) on top of 500 mL RPMI medium containing 3% FBS as chemoattractant. Inhibitors were added directly to the medium and mixed. Then the cells were incubated at 37 C in the CO 2 incubator for 16 hours. Non-invaded cells along with Matrigel in the upper chambers were scraped with a cotton tipped applicator and then the membranes were fixed in methanol, stained with 0.025% crystal violet, and observed under a Leica microscope at ×200.
Monga J., Valeriote F., Hwang C., Gadgeel S, & Ghosh J. (2023). Daclatasvir, an Antiviral Drug, Downregulates Tribbles 2 Pseudokinase and Resensitizes Enzalutamide-Resistant Prostate Cancer Cells. Molecular cancer therapeutics, 22(3).
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